A multiplex PCR assay for six Aedini species, including Aedes albopictus

Parasites & Vectors

Seong Tae Cho¹  Woo Jun Bang²  Jihun Ryu²  Min Hyeok Won²  Kwang Shik Choi³ 

[1] School of Life Sciences, Kyungpook National University, Biology building 226, Daehak-ro 80, Daegu, Korea;School of Life Sciences, Kyungpook National University, Biology building 226, Daehak-ro 80, Daegu, Korea;Research Institute for Dokdo and Ulleungdo Island, Kyungpook National University, Daegu, Korea;School of Life Sciences, Kyungpook National University, Biology building 226, Daehak-ro 80, Daegu, Korea;Research Institute for Dokdo and Ulleungdo Island, Kyungpook National University, Daegu, Korea;Research Institute for Phylogenomics and Evolution, Kyungpook National University, Daegu, Korea;

关键词: Aedini;    Aedes albopictus;    Internal transcribed spacer 2;    Multiplex PCR assay;   

DOI  :  10.1186/s13071-021-04871-7

来源: Springer

【 摘 要 】

BackgroundMosquitoes, as vectors of various human pathogens, are significant drivers of serious human illness. In particular, those species in the Aedini tribe, which typically transmit dengue virus, Chikungunya fever virus, and Zika virus, are increasing their range because of climate change and international commerce. In order to evaluate the risk of disease transmission, accurate mosquito species identification and monitoring are needed. The goal of this work was to develop a rapid and simple molecular diagnostic method for six morphologically similar Aedini species (Aedes flavopictus, Aedes albopictus, Ochlerotatus koreicus, Ochlerotatus japonicus, Ochlerotatus togoi and Ochlerotatus hatorii) in Korea.MethodsA total of 109 samples were assayed in this study. The internal transcribed spacer 2 (ITS2) regions from all six species were amplified, sequenced and analyzed using Mega 6. Following the identification of regions that were consistently different in terms of sequence between all six species, multiplex primers were designed to amplify these regions to generate species-specific fragments distinguishable by their size.ResultsUniquely sized fragments were generated in Ae. flavopictus (495 bp), Ae. albopictus (438 bp), Oc. koreicus (361 bp), Oc. togoi (283 bp), Oc. hatorii (220 bp) and Oc. japonicus (160 bp). Pairwise distance analysis showed that the difference was 35.0 ± 1.5% between Aedes spp. and Ochlerotatus spp., 17.4 ± 0.2% between Ae. albopictus and Ae. flavopictus and 11.1 ± 0.3% between Oc. koreicus and Oc. japonicus.ConclusionsIn this study, a multiplex PCR assay for six species of the Aedini tribe was developed. This assay is more accurate than morphological identification and will be useful for monitoring and controlling these vector mosquitoes.Graphical Abstract

【 授权许可】

CC BY